本文選題：干細(xì)胞 + 視網(wǎng)膜色素變性　； 參考：《第三軍醫(yī)大學(xué)學(xué)報(bào)》2017年11期

【摘要】：目的研究小鼠神經(jīng)干細(xì)胞(neural stem cells,NSCs)C17.2細(xì)胞系與從RCS大鼠變性視網(wǎng)膜中原代分離視網(wǎng)膜色素上皮細(xì)胞(retinal pigment epithelial,RPE)的線粒體交換方式及其對(duì)RPE細(xì)胞特性的影響。方法分離RCS大鼠RPE細(xì)胞并進(jìn)行培養(yǎng)和鑒定。分別用線粒體特異性標(biāo)記物Mitotracker-red和Mitotracker-green標(biāo)記RPE細(xì)胞和小鼠NSCs細(xì)胞的線粒體,將兩種細(xì)胞共培養(yǎng),激光共聚焦顯微鏡下觀察兩種細(xì)胞之間隧道納米管(tunneling nanotubes,TNT)的形成及線粒體轉(zhuǎn)運(yùn)方式。采用流式細(xì)胞儀檢測(cè)與NSCs共培養(yǎng)后RPE細(xì)胞的反應(yīng)性活性氧類(reactive oxygen species,ROS)水平、細(xì)胞周期和細(xì)胞凋亡水平的變化。結(jié)果來源于RCS大鼠視網(wǎng)膜的第3代RPE細(xì)胞生長(zhǎng)狀態(tài)良好,RPE65及Bestrophin蛋白陽(yáng)性率細(xì)胞均大于95%。與NSCs共培養(yǎng)24 h后,可見RPE細(xì)胞與NSCs間TNT的形成,NSCs中的線粒體向RPE細(xì)胞方向單向運(yùn)動(dòng),接受NSCs轉(zhuǎn)運(yùn)的線粒體后,RPE細(xì)胞的ROS水平降低(P0.01);RPE細(xì)胞的增殖能力增加,處于S期的細(xì)胞比例明顯增加(P0.01),細(xì)胞增殖指數(shù)(PI)顯著增加(P0.05);RPE細(xì)胞早期凋亡細(xì)胞比例顯著降低(P0.05)。結(jié)論 NSCs可通過與相鄰的變性RPE細(xì)胞之間形成TNT并向其轉(zhuǎn)運(yùn)線粒體而改善變性RPE細(xì)胞的存活。
[Abstract]:Objective to study the mitochondrial exchange patterns of neural stem cells from mouse neural stem cells and the primary isolation of retinal pigment epithelial cells (RPE) from the degenerative retina of RCS rats and their effects on the characteristics of RPE cells. Methods RCS rat RPE cells were isolated, cultured and identified. Mitochondrial specific markers (Mitotracker-red and Mitotracker-green) were used to label the mitochondria of RPE cells and mouse NSCs cells, respectively. The formation of tunneling nanotubes and mitochondrial transport patterns were observed under confocal laser microscope. The levels of reactive reactive oxygen species (Ros), cell cycle and apoptosis of RPE cells after co-culture with NSCs were detected by flow cytometry. Results the positive rates of RPE65 and Bestrophin protein in the third passage of RPE cells from the retina of RCS rats were higher than 95%. After co-cultured with NSCs for 24 h, the formation of TNT between RPE cells and NSCs cells showed that the mitochondria in RPE cells moved in one direction to RPE cells, and the ROS level of TNT cells after NSCs transport decreased the proliferation ability of P0.01RPE cells. The percentage of cells in S phase increased significantly, and the proliferation index (Pi) increased significantly. The percentage of early apoptosis cells in P0.05 RPE cells decreased significantly. Conclusion NSCs can improve the survival of denatured RPE cells by forming TNT between them and transporting mitochondria to them.
【作者單位】： 第三軍醫(yī)大學(xué)西南醫(yī)院全軍眼科中心 視覺損傷與再生修復(fù)重慶市重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展計(jì)劃(973計(jì)劃,2013CB967002)~~
【分類號(hào)】：R774.1

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